SSR Primers Development in Non-head ing Chinese Cabbage by ISSR-PCR and Streptavidin-coated Magnetic Beads Adsorption

ACTA HORTICULTURAE SINICA ›› 2006, Vol. 33 ›› Issue (1): 155-157.

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SSR Primers Development in Non-head ing Chinese Cabbage by ISSR-PCR and Streptavidin-coated Magnetic Beads Adsorption

Cui Xiumin¹;Hou Xilin^1*;and Dong Yuxiu²

(¹National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095,China; ²College of Life Science, Shandong Agricultural University, Tai'an 271018, China)Abstract:

Received:2005-03-07 Revised:2005-06-17 Online:2006-02-25 Published:2006-02-25

Abstract

Abstract: Two methods of ISSR-PCR and streptavidin-coated magnetic beads adsorption were used to isolate microsatellites from genomic DNA of non-heading Chinese cabbage (Brassica campestris ssp. chinensis) and a total of 41 SSR primers were successfully developed in this study. SSR primers ( IP₂ and IP₃)were designed on the base of the determined terminal sequence of nested PCR products in the ISSR-PCR method with the efficiency of 12% for SSR primer development. As to the method of streptavidin-coated magnetic beads adsorption, firstly the biotin-labeled SSR p robe was hybridized with digested genomic DNA and SSR fragments were enriched by absorption with strepavidin-coated magnetic beads and then theywere amp lified, cloned and
sequenced. On the base of the sequence flanking microsatellites SSR primers(P_L and P_R ) were designed with an efficiency of 9.6%.

Key words: Non-heading Chinese cabbage, Microsatellite, ISSR-PCR, Streptavidin-coated magnetic beads adsorption, SSR primer

Cui Xiumin;Hou Xilin;and Dong Yuxiu. SSR Primers Development in Non-head ing Chinese Cabbage by ISSR-PCR and Streptavidin-coated Magnetic Beads Adsorption[J]. ACTA HORTICULTURAE SINICA, 2006, 33(1): 155-157.

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SSR Primers Development in Non-head ing Chinese Cabbage by ISSR-PCR and Streptavidin-coated Magnetic Beads Adsorption

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